Bottom Line:
The process of formation of "grains" was dependent on the hemin concentration and incubation time.The possible mechanism of membrane nanostructure alterations is proposed.This research can be used to study the cell intoxication and analyze the action of various agents on RBC membranes.

ABSTRACTHemin is the product of hemoglobin oxidation. Some diseases may lead to a formation of hemin. The accumulation of hemin causes destruction of red blood cells (RBC) membranes. In this study the process of development of topological defects of RBC membranes within the size range from nanoscale to microscale levels is shown. The formation of the grain-like structures in the membrane ("grains") with typical sizes of 120-200 nm was experimentally shown. The process of formation of "grains" was dependent on the hemin concentration and incubation time. The possible mechanism of membrane nanostructure alterations is proposed. The kinetic equations of formation and transformation of small and medium topological defects were analyzed. This research can be used to study the cell intoxication and analyze the action of various agents on RBC membranes.

f2: The process of domains rising on the membrane under the hemin action. Comparison with the control.(a), (b),(c) For hemin concentration C = 0 (control). (d), (e),(f) For hemin concentration C = 1.1–1.2 mM. (a) and (d) The AFM 3D- images of RBCs 8 × 8 μm. (b) and (e) AFM 3D-images of membrane surfaces 1300 × 1300 nm (control) and 2400 × 2400 nm (surface with incipient domais). (c) and (f) Typical profiles of the surface in control membrane and in membrane with incipient domains. Interaction time of hemin with RBCs was 1 hour. The typical control cells were chosen out of the ensemble of 2820 cells, under the hemin action – out of 3010 cells. Membrane nanoscructure and profiles are represented as typical for given concentration and incubation time among 108 areas.

Mentions:
A representative AFM image (100 × 100 μm) of cells in the control smear is shown in Fig. 1a. Forty five images, each including 60–100 cells, were analyzed for each hemin concentration and each incubation time. Scanning of the fields 30 × 30 μm (Fig. 1 a–f) and 10 × 10 μm (Fig. 2, 3) was performed for more detailed AFM images.

f2: The process of domains rising on the membrane under the hemin action. Comparison with the control.(a), (b),(c) For hemin concentration C = 0 (control). (d), (e),(f) For hemin concentration C = 1.1–1.2 mM. (a) and (d) The AFM 3D- images of RBCs 8 × 8 μm. (b) and (e) AFM 3D-images of membrane surfaces 1300 × 1300 nm (control) and 2400 × 2400 nm (surface with incipient domais). (c) and (f) Typical profiles of the surface in control membrane and in membrane with incipient domains. Interaction time of hemin with RBCs was 1 hour. The typical control cells were chosen out of the ensemble of 2820 cells, under the hemin action – out of 3010 cells. Membrane nanoscructure and profiles are represented as typical for given concentration and incubation time among 108 areas.

Mentions:
A representative AFM image (100 × 100 μm) of cells in the control smear is shown in Fig. 1a. Forty five images, each including 60–100 cells, were analyzed for each hemin concentration and each incubation time. Scanning of the fields 30 × 30 μm (Fig. 1 a–f) and 10 × 10 μm (Fig. 2, 3) was performed for more detailed AFM images.

Bottom Line:
The process of formation of "grains" was dependent on the hemin concentration and incubation time.The possible mechanism of membrane nanostructure alterations is proposed.This research can be used to study the cell intoxication and analyze the action of various agents on RBC membranes.

ABSTRACTHemin is the product of hemoglobin oxidation. Some diseases may lead to a formation of hemin. The accumulation of hemin causes destruction of red blood cells (RBC) membranes. In this study the process of development of topological defects of RBC membranes within the size range from nanoscale to microscale levels is shown. The formation of the grain-like structures in the membrane ("grains") with typical sizes of 120-200 nm was experimentally shown. The process of formation of "grains" was dependent on the hemin concentration and incubation time. The possible mechanism of membrane nanostructure alterations is proposed. The kinetic equations of formation and transformation of small and medium topological defects were analyzed. This research can be used to study the cell intoxication and analyze the action of various agents on RBC membranes.